Expression of a fungal glucuronoyl esterase in Populus: effects on wood properties and saccharification efficiency.
Identifieur interne : 000310 ( Main/Corpus ); précédent : 000309; suivant : 000311Expression of a fungal glucuronoyl esterase in Populus: effects on wood properties and saccharification efficiency.
Auteurs : Madhavi Latha Gandla ; Marta Derba-Maceluch ; Xiaokun Liu ; Lorenz Gerber ; Emma R. Master ; Ewa J. Mellerowicz ; Leif J. JönssonSource :
- Phytochemistry [ 1873-3700 ] ; 2015.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Esterases.
- chemical , metabolism : Esterases, Glucuronic Acid, Polysaccharides.
- chemistry : Wood.
- enzymology : Phanerochaete.
- genetics : Phanerochaete, Populus.
- metabolism : Wood.
- Gene Expression, Hydrolysis.
Abstract
The secondary walls of angiosperms contain large amounts of glucuronoxylan that is thought to be covalently linked to lignin via ester bonds between 4-O-methyl-α-D-glucuronic acid (4-O-Me-GlcA) moieties in glucuronoxylan and alcohol groups in lignin. This linkage is proposed to be hydrolysed by glucuronoyl esterases (GCEs) secreted by wood-degrading fungi. We report effects of overexpression of a GCE from the white-rot basidiomycete Phanerochaete carnosa, PcGCE, in hybrid aspen (Populus tremula L. x tremuloides Michx.) on the wood composition and the saccharification efficiency. The recombinant enzyme, which was targeted to the plant cell wall using the signal peptide from hybrid aspen cellulase PttCel9B3, was constitutively expressed resulting in the appearance of GCE activity in protein extracts from developing wood. Diffuse reflectance FT-IR spectroscopy and pyrolysis-GC/MS analyses showed significant alternation in wood chemistry of transgenic plants including an increase in lignin content and S/G ratio, and a decrease in carbohydrate content. Sequential wood extractions confirmed a massive (+43%) increase of Klason lignin, which was accompanied by a ca. 5% decrease in cellulose, and ca. 20% decrease in wood extractives. Analysis of the monosaccharide composition using methanolysis showed a reduction of 4-O-Me-GlcA content without a change in Xyl contents in transgenic lines, suggesting that the covalent links between 4-O-Me-GlcA moieties and lignin protect these moieties from degradation. Enzymatic saccharification without pretreatment resulted in significant decreases of the yields of Gal, Glc, Xyl and Man in transgenic lines, consistent with their increased recalcitrance caused by the increased lignin content. In contrast, the enzymatic saccharification after acid pretreatment resulted in Glc yields similar to wild-type despite of their lower cellulose content. These data indicate that whereas PcGCE expression in hybrid aspen increases lignin deposition, the inhibitory effects of lignin are efficiently removed during acid pretreatment, and the extent of wood cellulose conversion during hydrolysis after acid pretreatment is improved in the transgenic lines possible due to reduced cell wall cross-links between cell wall biopolymers by PcGCE.
DOI: 10.1016/j.phytochem.2014.06.002
PubMed: 24997793
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Expression of a fungal glucuronoyl esterase in Populus: effects on wood properties and saccharification efficiency.</title><author><name sortKey="Latha Gandla, Madhavi" sort="Latha Gandla, Madhavi" uniqKey="Latha Gandla M" first="Madhavi" last="Latha Gandla">Madhavi Latha Gandla</name><affiliation><nlm:affiliation>Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Derba Maceluch, Marta" sort="Derba Maceluch, Marta" uniqKey="Derba Maceluch M" first="Marta" last="Derba-Maceluch">Marta Derba-Maceluch</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Liu, Xiaokun" sort="Liu, Xiaokun" uniqKey="Liu X" first="Xiaokun" last="Liu">Xiaokun Liu</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Gerber, Lorenz" sort="Gerber, Lorenz" uniqKey="Gerber L" first="Lorenz" last="Gerber">Lorenz Gerber</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Master, Emma R" sort="Master, Emma R" uniqKey="Master E" first="Emma R" last="Master">Emma R. Master</name><affiliation><nlm:affiliation>Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada.</nlm:affiliation></affiliation></author><author><name sortKey="Mellerowicz, Ewa J" sort="Mellerowicz, Ewa J" uniqKey="Mellerowicz E" first="Ewa J" last="Mellerowicz">Ewa J. Mellerowicz</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden. Electronic address: ewa.mellerowicz@slu.se.</nlm:affiliation></affiliation></author><author><name sortKey="Jonsson, Leif J" sort="Jonsson, Leif J" uniqKey="Jonsson L" first="Leif J" last="Jönsson">Leif J. Jönsson</name><affiliation><nlm:affiliation>Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden. Electronic address: leif.jonsson@chem.umu.se.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2015">2015</date><idno type="RBID">pubmed:24997793</idno><idno type="pmid">24997793</idno><idno type="doi">10.1016/j.phytochem.2014.06.002</idno><idno type="wicri:Area/Main/Corpus">000310</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000310</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Expression of a fungal glucuronoyl esterase in Populus: effects on wood properties and saccharification efficiency.</title><author><name sortKey="Latha Gandla, Madhavi" sort="Latha Gandla, Madhavi" uniqKey="Latha Gandla M" first="Madhavi" last="Latha Gandla">Madhavi Latha Gandla</name><affiliation><nlm:affiliation>Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Derba Maceluch, Marta" sort="Derba Maceluch, Marta" uniqKey="Derba Maceluch M" first="Marta" last="Derba-Maceluch">Marta Derba-Maceluch</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Liu, Xiaokun" sort="Liu, Xiaokun" uniqKey="Liu X" first="Xiaokun" last="Liu">Xiaokun Liu</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Gerber, Lorenz" sort="Gerber, Lorenz" uniqKey="Gerber L" first="Lorenz" last="Gerber">Lorenz Gerber</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</nlm:affiliation></affiliation></author><author><name sortKey="Master, Emma R" sort="Master, Emma R" uniqKey="Master E" first="Emma R" last="Master">Emma R. Master</name><affiliation><nlm:affiliation>Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada.</nlm:affiliation></affiliation></author><author><name sortKey="Mellerowicz, Ewa J" sort="Mellerowicz, Ewa J" uniqKey="Mellerowicz E" first="Ewa J" last="Mellerowicz">Ewa J. Mellerowicz</name><affiliation><nlm:affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden. Electronic address: ewa.mellerowicz@slu.se.</nlm:affiliation></affiliation></author><author><name sortKey="Jonsson, Leif J" sort="Jonsson, Leif J" uniqKey="Jonsson L" first="Leif J" last="Jönsson">Leif J. Jönsson</name><affiliation><nlm:affiliation>Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden. Electronic address: leif.jonsson@chem.umu.se.</nlm:affiliation></affiliation></author></analytic><series><title level="j">Phytochemistry</title><idno type="eISSN">1873-3700</idno><imprint><date when="2015" type="published">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Esterases (genetics)</term><term>Esterases (metabolism)</term><term>Gene Expression (MeSH)</term><term>Glucuronic Acid (metabolism)</term><term>Hydrolysis (MeSH)</term><term>Phanerochaete (enzymology)</term><term>Phanerochaete (genetics)</term><term>Polysaccharides (metabolism)</term><term>Populus (genetics)</term><term>Wood (chemistry)</term><term>Wood (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Esterases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Esterases</term><term>Glucuronic Acid</term><term>Polysaccharides</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Wood</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Phanerochaete</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Wood</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Expression</term><term>Hydrolysis</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The secondary walls of angiosperms contain large amounts of glucuronoxylan that is thought to be covalently linked to lignin via ester bonds between 4-O-methyl-α-D-glucuronic acid (4-O-Me-GlcA) moieties in glucuronoxylan and alcohol groups in lignin. This linkage is proposed to be hydrolysed by glucuronoyl esterases (GCEs) secreted by wood-degrading fungi. We report effects of overexpression of a GCE from the white-rot basidiomycete Phanerochaete carnosa, PcGCE, in hybrid aspen (Populus tremula L. x tremuloides Michx.) on the wood composition and the saccharification efficiency. The recombinant enzyme, which was targeted to the plant cell wall using the signal peptide from hybrid aspen cellulase PttCel9B3, was constitutively expressed resulting in the appearance of GCE activity in protein extracts from developing wood. Diffuse reflectance FT-IR spectroscopy and pyrolysis-GC/MS analyses showed significant alternation in wood chemistry of transgenic plants including an increase in lignin content and S/G ratio, and a decrease in carbohydrate content. Sequential wood extractions confirmed a massive (+43%) increase of Klason lignin, which was accompanied by a ca. 5% decrease in cellulose, and ca. 20% decrease in wood extractives. Analysis of the monosaccharide composition using methanolysis showed a reduction of 4-O-Me-GlcA content without a change in Xyl contents in transgenic lines, suggesting that the covalent links between 4-O-Me-GlcA moieties and lignin protect these moieties from degradation. Enzymatic saccharification without pretreatment resulted in significant decreases of the yields of Gal, Glc, Xyl and Man in transgenic lines, consistent with their increased recalcitrance caused by the increased lignin content. In contrast, the enzymatic saccharification after acid pretreatment resulted in Glc yields similar to wild-type despite of their lower cellulose content. These data indicate that whereas PcGCE expression in hybrid aspen increases lignin deposition, the inhibitory effects of lignin are efficiently removed during acid pretreatment, and the extent of wood cellulose conversion during hydrolysis after acid pretreatment is improved in the transgenic lines possible due to reduced cell wall cross-links between cell wall biopolymers by PcGCE.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24997793</PMID><DateCompleted><Year>2015</Year><Month>12</Month><Day>21</Day></DateCompleted><DateRevised><Year>2015</Year><Month>03</Month><Day>27</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1873-3700</ISSN><JournalIssue CitedMedium="Internet"><Volume>112</Volume><PubDate><Year>2015</Year><Month>Apr</Month></PubDate></JournalIssue><Title>Phytochemistry</Title><ISOAbbreviation>Phytochemistry</ISOAbbreviation></Journal><ArticleTitle>Expression of a fungal glucuronoyl esterase in Populus: effects on wood properties and saccharification efficiency.</ArticleTitle><Pagination><MedlinePgn>210-20</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.phytochem.2014.06.002</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S0031-9422(14)00253-2</ELocationID><Abstract><AbstractText>The secondary walls of angiosperms contain large amounts of glucuronoxylan that is thought to be covalently linked to lignin via ester bonds between 4-O-methyl-α-D-glucuronic acid (4-O-Me-GlcA) moieties in glucuronoxylan and alcohol groups in lignin. This linkage is proposed to be hydrolysed by glucuronoyl esterases (GCEs) secreted by wood-degrading fungi. We report effects of overexpression of a GCE from the white-rot basidiomycete Phanerochaete carnosa, PcGCE, in hybrid aspen (Populus tremula L. x tremuloides Michx.) on the wood composition and the saccharification efficiency. The recombinant enzyme, which was targeted to the plant cell wall using the signal peptide from hybrid aspen cellulase PttCel9B3, was constitutively expressed resulting in the appearance of GCE activity in protein extracts from developing wood. Diffuse reflectance FT-IR spectroscopy and pyrolysis-GC/MS analyses showed significant alternation in wood chemistry of transgenic plants including an increase in lignin content and S/G ratio, and a decrease in carbohydrate content. Sequential wood extractions confirmed a massive (+43%) increase of Klason lignin, which was accompanied by a ca. 5% decrease in cellulose, and ca. 20% decrease in wood extractives. Analysis of the monosaccharide composition using methanolysis showed a reduction of 4-O-Me-GlcA content without a change in Xyl contents in transgenic lines, suggesting that the covalent links between 4-O-Me-GlcA moieties and lignin protect these moieties from degradation. Enzymatic saccharification without pretreatment resulted in significant decreases of the yields of Gal, Glc, Xyl and Man in transgenic lines, consistent with their increased recalcitrance caused by the increased lignin content. In contrast, the enzymatic saccharification after acid pretreatment resulted in Glc yields similar to wild-type despite of their lower cellulose content. These data indicate that whereas PcGCE expression in hybrid aspen increases lignin deposition, the inhibitory effects of lignin are efficiently removed during acid pretreatment, and the extent of wood cellulose conversion during hydrolysis after acid pretreatment is improved in the transgenic lines possible due to reduced cell wall cross-links between cell wall biopolymers by PcGCE.</AbstractText><CopyrightInformation>Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Latha Gandla</LastName><ForeName>Madhavi</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Derba-Maceluch</LastName><ForeName>Marta</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Xiaokun</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gerber</LastName><ForeName>Lorenz</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Master</LastName><ForeName>Emma R</ForeName><Initials>ER</Initials><AffiliationInfo><Affiliation>Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Mellerowicz</LastName><ForeName>Ewa J</ForeName><Initials>EJ</Initials><AffiliationInfo><Affiliation>Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden. Electronic address: ewa.mellerowicz@slu.se.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jönsson</LastName><ForeName>Leif J</ForeName><Initials>LJ</Initials><AffiliationInfo><Affiliation>Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden. Electronic address: leif.jonsson@chem.umu.se.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2014</Year><Month>07</Month><Day>02</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Phytochemistry</MedlineTA><NlmUniqueID>0151434</NlmUniqueID><ISSNLinking>0031-9422</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011134">Polysaccharides</NameOfSubstance></Chemical><Chemical><RegistryNumber>8A5D83Q4RW</RegistryNumber><NameOfSubstance UI="D020723">Glucuronic Acid</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.1.-</RegistryNumber><NameOfSubstance UI="D004950">Esterases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D004950" MajorTopicYN="N">Esterases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015870" MajorTopicYN="N">Gene Expression</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020723" MajorTopicYN="N">Glucuronic Acid</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006868" MajorTopicYN="N">Hydrolysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020075" MajorTopicYN="N">Phanerochaete</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011134" MajorTopicYN="N">Polysaccharides</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D032107" MajorTopicYN="N">Populus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014934" MajorTopicYN="N">Wood</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">CE15</Keyword><Keyword MajorTopicYN="N">Enzymatic saccharification</Keyword><Keyword MajorTopicYN="N">Glucuronoyl esterase</Keyword><Keyword MajorTopicYN="N">Hybrid aspen</Keyword><Keyword MajorTopicYN="N">Populus</Keyword><Keyword MajorTopicYN="N">Secondary cell wall</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2014</Year><Month>02</Month><Day>10</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2014</Year><Month>04</Month><Day>28</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2014</Year><Month>06</Month><Day>05</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>7</Month><Day>7</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2014</Year><Month>7</Month><Day>7</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2015</Year><Month>12</Month><Day>22</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">24997793</ArticleId><ArticleId IdType="pii">S0031-9422(14)00253-2</ArticleId><ArticleId IdType="doi">10.1016/j.phytochem.2014.06.002</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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